Team:UC Davis/notebook/initiator.html

From 2010.igem.org

(Difference between revisions)
Line 81: Line 81:
<ul>
<ul>
-
<li><a href="https://2010.igem.org/Team:UC_Davis/protocols/ligation.html" class="help">Ligate</a> the RBS and BBa_C0051 fragments together and <a href="https://2010.igem.org/Team:UC_Davis/protocols/transformation.html" class="help">transform</a> into competent cells.</li>
+
<li><a href="https://2010.igem.org/Team:UC_Davis/protocols/ligation.html" class="help">Ligate</a> the RBS and BBa_C0051 fragments together and <a href="https://2010.igem.org/Team:UC_Davis/protocols/transformation.html" class="help">transform</a> on carb plates.</li>
 +
<li>Depending on the outcome of visible colonies on the experimental and control plates, a <a href="https://2010.igem.org/Team:UC_Davis/protocols/pcrscreen.html" class="help">PCR screening</a> may be needed.
</ul><br />
</ul><br />
Line 89: Line 90:
<ul>
<ul>
<li>It is most practical to cut RBS as a vector because RBS is less than 100 bp long as an insert, making it hard to see and extract on the gel.</li>
<li>It is most practical to cut RBS as a vector because RBS is less than 100 bp long as an insert, making it hard to see and extract on the gel.</li>
-
<li>The sticky ends from the XbaI and SpeI, when ligated together, will form an 8 bp scar.</li>
+
<li>The sticky ends from the XbaI and SpeI, when ligated together, will form an 8 bp scar.  Thus, XbaI and SpeI will no longer be able to cut here.</li>
</ul>
</ul>
                            
                            

Revision as of 09:56, 19 September 2010

The Initiator

Starting Up

  • Hydrated parts: BBa_B0034, BBa_C0051, BBa_B0015, BBa_R0082.
  • Transformed these parts into DH5α competent cells.


RBS to BBa_C0051


  • Ligate the RBS and BBa_C0051 fragments together and transform on carb plates.
  • Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.


Assembly notes
  • It is most practical to cut RBS as a vector because RBS is less than 100 bp long as an insert, making it hard to see and extract on the gel.
  • The sticky ends from the XbaI and SpeI, when ligated together, will form an 8 bp scar. Thus, XbaI and SpeI will no longer be able to cut here.
We would like to take a moment to thank all of our sponsors for their very generous donations, as we could not have done this without your help!

We would also like to thank and acknowledge:
Our Advisors
Marc Facciotti
Ilias Tagkopoulos
Technical Guidance
David Larsen
Andrew Yao
Visiting iGEMer
Jia Li of Zhejiang University (TEAM ZJU-China)
cI Promoter Screen
Drew Endy - Stanford
Thomas Schneider - NIH
Want to sponsor us? Send an email to mtfacciotti@ucdavis.edu to discuss various ways you can help! :)

Retrieved from "http://2010.igem.org/Team:UC_Davis/notebook/initiator.html"